8. タバコ培養細胞の液体窒素保存(平成5年度第39回凍結及び乾燥研究会研究報告)

概要

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Cell suspension of Nicotiana tabacum L. cv. BY2 was cultured with a modified LS medium. The cells were examined to vitrify through two methods. The 7-days culture was replaced in the medium with 1M sucrose and cultured for 1 day. In the method 1, the cells were vitrified with PVS2 (by SAKAI) in liquid nitrogen. In the method 2, the cells were transferred to the medium containing 2M glycerol plus 0.4M sorbitol, and then to that containing 4.5M glycerol plus 1.46M sorbitol successively. Cells in the concentrated medium were vitrified similarly to the method 1. Plasmolysis of the cells was observed in both methods, but more mildly in the method 2. The ratio of heat of devitrification to that of thawing was 0.82 or 0.67 in the two methods, respectively. The thawed cells in both methods were hard to grow on the solid medium. Only the growth was observed on a solid medium through nursing culture plate of the cells from methed 2.
低温生物工学会の論文
1993-10-20